mTORC2 Activation Mediated by Mesenchymal Stem Cell-Secreted Hepatocyte Growth Factors for the Recovery of Lipopolysaccharide-Induced Vascular Endothelial Barrier

Figure 5

Effects of mTORC2/Akt and mTORC2/PKC signaling to HGF on endothelial barrier in LPS-induced PMVECs. PMVECs were treated with HGF (20 ng/ml), with or without stimulation with LPS (100 ng/ml) for 24 h. And enzastaurin (2 μM) and AZD5363 (1 μM) were, respectively, used as PKC and Akt inhibitors. PtdIns(3,4,5)P3 (25ng/ml) was used to active mTORC2 as positive control. (a) Flow cytometry scatter plot of 4 h VE-cadherin expression with HGF in LPS-induced PMVECs under Akt and PKC inhibition. (b) Flow cytometry cell counts (%) of 4 h VE-cadherin expression with HGF in LPS-induced PMVECs under Akt and PKC inhibition. (c) The effects of Akt and PKC inhibitors to relative paracellular permeability of HGF on LPS-induced PMVEC permeability with Alexa Fluor 647-dextran. (d) The effects of Akt and PKC inhibitors to relative transcellular permeability of HGF on LPS-induced PMVEC permeability with Alexa Fluor 647-BSA. Results are (). . (e) Evaluation of HGF on the mTORC signaling pathway for western blot.

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